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Biotechnology is an ever-evolving field of science that is instrumental in improving quality of human life, particularly in the medical arena. Optogenetics, an area of biotechnology, involves genetic modification of cells to express light-sensitive ion channels, which allows for use of light to manipulate behavior. This educational module utilizes an approach to bridge optogenetics, cellular metabolism, and animal behavior for student-driven inquiry in college courses. Drosophila melanogaster larvae that have been modified to express Channelrhodopsin-2 (ChR2) in motor neurons serve as models for this module. Students are able to connect temperature, metabolic rate, and gene expression through data collection of behavioral responses to light stimuli exhibited by larvae stored in various temperatures. Students are also able to observe the role of cofactors in metabolic processes via larvae that have been fed all-trans retinal (ATR), which is a cofactor to ChR2. Students will analyze and interpret data in order to make a claim about how the two variables investigated (temperature and cofactor) impact the behavior of the target organism. The final component of the module allows students to form connections between the application of neuroscience as exhibited in the module to trends in scientific publications related to optogenetics. This module is presented in a variety of ways to be readily modified depending on equipment available in the classroom and the academic level of students. The modules were designed to be used in a hybrid or remote format with data provided online with sample data sets as well as movies of larval behaviors. The exercises can also be conducted by in class participants.

ABLE Poster

ABLE paper- draft

Movie to go with poster MP4

Protocol for class: MS word and poster

Background reading in optogenetics and fly behaviors:

Lights and Larvae: Using optogenetics to teach recombinant DNA and neurobiology. The Science Teacher 81 (#6):2-9. [PDF] .cover

Optogenetics in the teaching laboratory: [PDF]

Optogenetic manipulation of neural circuits and behavior in Drosophila larvae [PDF]

Channelrhodopsin reveals experience-dependent influences on courtship [PDF]

Selective neural activity in multiple freely moving Drosophila adults [PDF]

Nature 2015- review on optogenetics [PDF]

Nature 2015- review on optogenetics- commentary [PDF]

Nature 2015- review on optogenetics- neuroscience [PDF]

Optopharmacological tools for restoring visual function in degenerative retinal diseases (link to abstract)

Top 10 discoveries in 10 years of optogenetics (link to)

Ion channels with ChR channels (PDF)

http://www.jove.com/video/50513/optogenetic-perturbation-neural-activity-with-laser-illumination-semi

Interesting side articles and links related to this topic

http://www.sciencedaily.com/releases/

Anatomical and genotype-specific mechanosensory responses in Drosophila melanogaster larvae. Neuroscience Research 83:54-63 [PDF]

Table of which lines activate which type of neurons (PDF, MS Word )

Sample movies of larval behavior: (hot links with names-raw data files MP4)

(Incubator = 27C for 24 hours 2nd instar to 3rd instar)

CHR2XXL x 24B (body wall muscles)
Room temp/No ATR larva 1
Room temp/No ATR larva 2
Room temp/No ATR larva 3
Room temp/ATR larva 1 (preps1-3)
Room temp/ATR larva 2 (preps1-3)
Room temp/ATR larva 3 (preps1-3)
Incubator/No ATR larva 1....(Preps1 & 2 and a parental Chr2XXL)
Incubator/No ATR larva 2....(Preps1 & 2 and a parental Chr2XXL)
Incubator/No ATR larva 3
Incubator /ATR larva 1
Incubator /ATR larva 2
Incubator /ATR larva 3

CHR2XXL x D42 (Motor neurons)
Room temp/No ATR larva 1
Room temp/No ATR larva 2
Room temp/No ATR larva 3
Room temp/ATR larva 1
Room temp/ATR larva 2
Room temp/ATR larva 3
Incubator/No ATR larva 1
Incubator/No ATR larva 2
Incubator/No ATR larva 3
Incubator /ATR larva 1
Incubator /ATR larva 2....(preps 2 & 3)
Incubator /ATR larva 3....(preps 2 & 3)

Controls (Chr2 only-parental line):

CONTROL-No ATR-20C-Chr2 only no cross 5 larvae dish 1
CONTROL-No ATR-Incubator-Chr2 only no cross 5 larvae dish 1
CONTROL-ATR-20C-Chr2 only no cross 6 larvae dish 1
CONTROL ATR Incubator Chr2 only no cross 5larvae dish 1

YOUTUBE links

Controls (Chr2 only-parental line):

CONTROL-No ATR-20C-Chr2 only no cross 5 larvae dish 1 https://youtu.be/GmyVFbbB9i0
CONTROL-No ATR-Incubator-Chr2 only no cross 5 larvae dish 1 https://youtu.be/PzzQZ03BH1g
CONTROL-ATR-20C-Chr2 only no cross 6 larvae dish 1 https://youtu.be/rFa03rk0jCk
CONTROL ATR Incubator Chr2 only no cross 5larvae dish 1 https://youtu.be/EtPU2JILNXQ

Participants designing this content are :

Tawny Aguayo-Williams,West Jessamine High School, 2101 Wilmore Rd, Nicholasville, KY 40356.

Vaaragie Subramaniam, Department of Biology, Univ. of Kentucky, Lexington, KY. 40506-0225

Robin L. Cooper, Department of Biology, Univ. of Kentucky, Lexington, KY. 40506-0225

Doug Harrison, Department of Biology, Univ. of Kentucky, Lexington, KY. 40506-0225

Brett Criswell, Department of Secondary Education, West Chester Univ., West Chester, PA. 19383